1 Introduction to forensicgenetics

The development and application of genetics has revolutionized forensic science. In1984, the analysis of polymorphic regions of DNA produced what was termed ‘aDNA fingerprint’ [1]. The following year, at the request of the United KingdomHome Office, DNA profiling was successfully applied to casework when it was usedto resolve an immigration dispute [2]. In 1986, DNA evidence was used for the firsttime in a criminal case involving the murder of two young women in Leicestershire,UK: DNA analysis exonerated one individual who had confessed to one of themurders, and following a mass screen of approximately 5000 individuals, identifiedColin Pitchfork as the murderer. He was convicted in January 1988 [3].1

Following on from early success in both civil and criminal cases, the use ofgenetics was rapidly adopted by the forensic community and now plays an importantrole worldwide in both the investigation of crime and in relationship testing. Thescope and scale of DNA analysis in forensic science is set to continue expanding forthe foreseeable future.

Forensic genetics

The work of the forensic geneticist will vary widely depending on the laboratoryand country that they work in, and can involve the analysis of material recoveredfrom a scene of crime, kinship testing and the identification of human remains.In some cases, it can even be used for the analysis of DNA from plants [6–18];animals [19–36] including insects [37–60]; and microorganisms [61–67]. The focusof this book is the analysis of biological material that is recovered from the sceneof crime – this is central to the work of most forensic laboratories. Kinship testingwill be dealt with separately in Chapter 11 and a brief introduction is given to thetesting of non-human material in Chapter 14.

Forensic laboratories receive material that has been recovered from scenes ofcrime, and reference samples from both suspects and victims. The role of forensicgenetics within the investigative process is to compare samples recovered from crime

1 Note: The first convictions in criminal cases came in 1987, when DNA evidence played an important role in the convictionof two rapists: Robert Melias in the UK and Tommy Lee Andrews in the USA [4, 5].

An Introduction to Forensic Genetics Second Edition William Goodwin, Adrian Linacre and Sibte Hadi© 2011 John Wiley & Sons, Ltd

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HTED M

ATERIAL

2 INTRODUCTION TO FORENSIC GENETICS

Forensic biologist

Referencesamples

from suspects Forensic geneticist

Report/intelligence

Referencesamples

from victims

Samples recovered fromscenes of crime

Figure 1.1 The role of the forensic geneticist is to assess whether samples recovered from a crimescene match to a suspect. Reference samples are provided from suspects and also victims of crime

scenes with suspects and possibly victims, resulting in a report that can be presentedin court or intelligence that may inform an investigation (Figure 1.1).

Several stages are involved in the analysis of genetic evidence (Figure 1.2) andeach of these is covered in detail in the following chapters. In some organizationsone person will be responsible for collecting the evidence, the biological and geneticanalysis of samples and ultimately presenting the results to a court of law. However,the trend in many larger organizations is for individuals to be responsible for onlya very specific task within the process, such as the extraction of DNA from theevidential material or the analysis and interpretation of DNA profiles that have beengenerated by other scientists, or just reporting the findings.

A brief history of forensic genetics

In 1900 Karl Landsteiner described the ABO blood grouping system and observedthat individuals could be placed into different groups based on their blood type. Thiswas the first step in the development of forensic haemogenetics. Following on fromthis, numerous blood group markers and soluble blood serum protein markers werecharacterized and could be analysed in combination to produce highly discriminatoryprofiles. The serological techniques were a powerful tool but were limited in manyforensic cases by the amount of biological material that was required to providehighly discriminating results. Proteins are also prone to degradation on exposure tothe environment.

In the 1960s and 1970s, developments in molecular biology, including restrictionenzymes, Sanger sequencing [68] and Southern blotting [69], enabled scientists toexamine DNA sequences. By 1978, DNA polymorphisms could be detected using

A BRIEF HISTORY OF FORENSIC GENETICS 3

Identification/Collection of material

DNA extraction

Quantification of DNA

PCR amplification

Detection of PCR products(DNA Profile)

Analysis and interpretation of profile

Statistical evaluation of DNA profile

Report

Characterization of material

Event

Transfer of material

Figure 1.2 Processes involved in generating a DNA profile following a crime. Some types ofmaterial, in particular blood and semen, are often characterized before DNA is extracted, allowingthe DNA evidence to be linked to a cell type and possible transfer mechanism

Southern blotting [70] and in 1980 the analysis of the first highly polymorphic locuswas reported, where the polymorphism was caused by differences in the lengths ofthe alleles [71].

It was not until September 1984 that Alec Jeffreys realized the potential forensicapplication of the minisatellite loci [1, 72, 73]. The technique developed by Jeffreysentailed extracting DNA and cutting it with a restriction enzyme before carrying outagarose gel electrophoresis, Southern blotting and probe hybridization to detect thepolymorphic loci. The end result was a series of black bands on X-ray film, whichwas called a DNA fingerprint (Figure 1.3).

4 INTRODUCTION TO FORENSIC GENETICS

(a) (b)

C M FM TC UD1 UD2 UD3

Figure 1.3 (a) The first ever DNA fingerprint, produced in Alec Jeffreys’ laboratory on the 10th

September 1984. It shows the banding pattern from a mother, child, and father: the bands in thechild’s profile can be attributed to either the mother or the father. (b) Profiles from a mother, atested child (TC) (Immigration Officials in the UK did not believe that the boy was the biological sonof the mother) and three of the mother’s undisputed children (UD 1-3): the results demonstratedthat the tested child was indeed the biological son of the woman and was therefore allowed tostay in the UK [2]. (Images provided by Sir Prof Alec Jeffreys, Department of Genetics, Universityof Leicester, UK)

With the first DNA fingerprints the multi-locus probes (MLPs) detected severalminisatellite loci simultaneously, leading to the multiple band patterns. While themulti-banded fingerprints were highly informative they were difficult to interpret.New probes were designed that were specific to one locus (single locus probes, SLPs)and therefore produced only one or two bands for each individual [74] (Figure 1.4).

Minisatellite analysis was a powerful tool but suffered from several limitations: arelatively large amount of DNA was required; it would not work with degraded DNA;comparison between laboratories was difficult; and the analysis was time consuming.Even so, the use of minisatellite analysis, using SLPs, was common for several years[75] until it was replaced by polymerase chain reaction (PCR)-based systems.

A critical development in the history of forensic genetics came with the adventof a process that can amplify specific regions of DNA – the PCR (see Chapter 5).The PCR process was conceptualized in 1983 by Kary Mullis, a chemist workingfor the Cetus Corporation in the USA [76]. The development of PCR has had aprofound effect on all aspects of molecular biology including forensic genetics, andin recognition of the significance of the development of the technique Kary Mulliswas awarded the Nobel Prize for Chemistry in 1993.

A BRIEF HISTORY OF FORENSIC GENETICS 5

Ladder Ladder

K562 K562

Ladder

Figure 1.4 Minisatellite analysis using a single locus probe: ladders were run alongside the testedsamples that allowed the size of the DNA fragments to be estimated. A control sample labelledK562 was analysed along with the tested samples

The PCR increases the sensitivity of DNA analysis to the point where DNA profilescan be generated from just a few cells, reduces the time required to produce a profile,can be used with degraded DNA and allows just about any polymorphism in thegenome to be analysed.

The first application of PCR in a forensic case involved the analysis of singlenucleotide polymorphisms in the human leukocyte antigen (HLA)-DQα locus (partof the major histocompatibility complex (MHC)) [77] (see Chapter 12). This wassoon followed by the analysis of short tandem repeats (STRs), which are currentlythe most commonly used genetic markers in forensic science (see Chapters 6–8).The rapid development of technology for analysing DNA includes advances in DNAextraction and quantification methodology, the development of commercial PCR-based typing kits and equipment for detecting DNA polymorphisms.

In addition to technical advances, another important part of the development ofDNA profiling that has had an impact on the whole field of forensic science isquality assurance. The admissibility of DNA evidence was seriously challenged in theUSA in 1989 – People v. Castro [78]; this and subsequent cases in many countrieshave resulted in increased levels of standardization and quality assurance in forensicgenetics and other areas of forensic science. As a result, the accreditation of bothlaboratories and individuals is an increasingly important issue in forensic science.The combination of technical advances, high levels of standardization and quality

6 INTRODUCTION TO FORENSIC GENETICS

assurance have led to forensic DNA analysis being recognized as a robust and reliableforensic tool.

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